The goal of this research project is to explore the structural heterogeneity of amyloid aggregates and the relationships of this conformational variation to the toxicity or pathogenic activities of amyloid oligomers. Several distinct types of amyloid deposits accumulate in disease brain and current evidence suggests that soluble, oligomeric forms of Ass may play primary role in pathogenesis. Recent results indicate that conformation-dependent monoclonal antibodies can distinguish between different types soluble Ass oligomers. These antibodies also distinguish other conformations of Ass, including monomers, fibrils and natively-folded APP. We have recently prepared two additional conformation-dependent antisera that recognize generic epitopes associated with amyloid fibrils and pore-like annular protofibrils that are formed from many different types of amyloids (see Preliminary Data, below). These antibodies are complementary to the anti-oligomer antibody, A11 and recognize epitopes that are specific to fibrils and annular protofibril aggregates. We hypothesize that these conformationally distinct assembly states of Ass are differentially associated with AD pathogenesis. We anticipate that these results will help clarify some apparent inconsistencies and conflicting data, such as the observations that the total Ass amyloid deposited correlates poorly with disease and some people have large amounts of amyloid and are cognitive normal, while other brain samples that have little observable amyloid deposits are associated with cognitive dysfunction. We hypothesize that the distinct types of soluble oligomeric or annular protofibril forms of Ass have distinct toxicities. Conformation-dependent antibodies hold the potential of identifying and distinguishing specific assembly states because they only recognize a specific misfolded state and do not react with the natively folded precursor protein. The specific aims of this project will address the following questions: What is the conformational diversity of amyloid aggregates? What is the structural basis of conformation dependent antibody specificity and amyloid oligomer structural? What are the relationships between different conformational states of amyloids? What is the pathological significance of the different amyloid conformational states? The answers to these questions should provide insight into the range of amyloid oligomer conformational diversity and monoclonal antibody reagents that distinguish different conformations of oligomers. This may provide a more rational structural basis for the classification of oligomers and provide insight into the variability in oligomer preparations reported by different laboratories. Determining the 3 dimensional structure of the monoclonal Fabs bound to amyloid oligomers may provide unprecedented insight into the structure of amyloid oligomers and the mechanism of specific antibody recognition that may be useful for development of immunological therapeutics that target oligomer formation or prevent their interaction with cellular targets. The identification of small molecules that specifically inhibit the formation of different types of oligomers should help to clarify whether the oligomers are intermediates in the formation of higher order structures, like fibrils or whether they represent stable alternative end products. This aim may also provide small molecule lead compounds that specifically inhibit amyloid oligomer formation for therapeutic development. The characterization of which types of amyloid oligomers are more closely related to pathogenesis in human and transgenic mouse brain may help to identify targets for therapeutic development. PHS 398/2590 (Rev. 04/06) Page Continuation Format Page PUBLIC HEALTH RELEVANCE: Amyloid aggregates are believed to play a central role in the development of neurodegenerative diseases, like Alzheimer, Parkinson, Huntington and prion diseases. The goal of this proposal is to determine how many different types of amyloid aggregates exist and to determine the precise 3 dimensional structure of the amyloid aggregates. The proposal will also examine which of these aggregates is more toxic to neurons and most closely related to disease. An additional benefit is that this proposal may produce monoclonal antibodies and drugs that specifically target these amyloid aggregates for therapeutic development.